Expression and activation of the epidermal growth factor receptor in differentiating cells of the developing and post-hatching chicken lens.

The epidermal growth factor receptor is hypothesized to play an important role in the post-natal growth and differentiation of the ocular lens. Immunohistochemistry and western blotting were utilized to examine the distribution and activation of the epidermal growth factor receptor in embryonic and post-hatching chicken lenses. Although present at constant levels within epithelial cells throughout embryonic development, the receptor becomes increasingly activated on a highly conserved tyrosine residue necessary for intracellular signal transduction as hatching approaches. After hatching, activated receptors are found in epithelial cells committed to fiber cell formation and in fiber cells undergoing initial stages of terminal differentiation. Activated receptors could not be identified in central epithelial cells or nuclear fiber cells. This pattern persists until at least one year post-hatching. These data indicate that the epidermal growth factor receptor is positioned to influence not only post-natal patterns of lenticular gene expression but also the greatest amount of lens growth and development.

Thymosin beta 4 stimulates laminin-5 production independent of TGF-beta.

Thymosin beta 4 (Tbeta(4)) stimulates epithelial cell migration and promotes laminin-5 (LM-5) expression. Using gene expression analysis with human corneal epithelial cells treated with Tbeta(4), we find that both LM-5 gamma2 chain and transforming growth factor beta 1 (TGFbeta-1) are increased by more than 2-fold over untreated cells. These findings were confirmed by RT-PCR and at the protein level. Although TGFbeta-1 increases LM-5 synthesis in a dose-dependent manner, it does not appear to be the mechanism by which Tbeta(4) acts on LM-5 gamma2 chain synthesis based on three independent experiments. In a time-course analysis, Tbeta(4) increases LM-5 gamma2 chain expression at 2 h and peaks at 6 h, while TGFbeta-1 increases LM-5 gamma2 chain expression only at 4 h and peaks at 8 h. When Tbeta(4)-induced LM-5 gamma2 chain expression is blocked with neutralizing antibodies to TGFbeta-1, LM-5 gamma2 chain expression is increased. Finally, in TGFbeta-1 knock-out mice, Tbeta(4) increases LM-5 gamma2 chain expression to levels higher than that observed in wild-type mice treated with Tbeta(4). These findings demonstrate that Tbeta(4) induces both TGFbeta-1 and LM-5 gamma2 chain expression in corneal epithelial cells. Tbeta(4) and TGFbeta-1 increase LM-5 gamma2 chain expression by independent pathways. Suppression of TGFbeta-1 further increases LM-5 gamma2 chain expression.